HSP For Use in Treatment for Imiquimod Related Side Effects

ABSTRACT

The invention relates to a healthcare product comprising (i) a component selected from the group of heat shock proteins from alfalfa and heat shock protein hydrolysates from alfalfa, the product further comprising imiquimod 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt or derivative thereof. 
     Further, the invention relates to a healthcare product for use in the prophylactic or therapeutic treatment of a skin disorder. Further, the invention relates to HSP for use for use in preventing the occurrence of a negative-side effect of a treatment with imiquimod, or alleviating such side effect.

THE INVENTION

The invention relates to a healthcare product comprising apharmaceutical compound for treating a skin disorder. The inventionfurther relates to a heat shock protein (Hsp) for medical use.

Various healthcare products for treating the skin are known. Theproducts may be purely cosmetic compositions, essentially cosmeticcompositions with medical benefits, over the counter drugs orprescription drugs.

Healthcare products, such as skincare products, comprising apharmaceutically compound can be unsatisfactory in that the effect ofthe treatment is less than desired (e.g. low effect or slow effect inrealising the intended purpose, for instance, wound healing, treatmentof infection or inflammation) or in that the pharmaceutical compound maycause an unwanted side-effect. In particular skincare productscomprising a pharmaceutical compound can have a significant risk ofcausing skin irritation, itchiness, skin redness (erythema), scaling ofthe skin, or swelling of the skin.

It is an object of the invention to provide an alternative healthcareproduct, in particular a skincare product or an Hsp containing agent,suitable for use in the prevention or therapeutic treatment of a skindisorder. In particular it is an object to provide such product, thathas satisfactory efficacy and wherein the risk of a side-effect to theskin is reduced at least for some subjects, or wherein theside-effect—if it manifests itself—is less severe or vanished within arelatively short time-span, at least for some subjects.

The inventors have found that Hsp used in combination with a specificpharmaceutical is advantageous in that the medical effect of thepharmaceutical is improved and/or in that a negative side-effect of thepharmaceutical is alleviated or the manifestation of the side-effect isavoided.

Accordingly, the present invention relates to a healthcare productcomprising (i) a component selected from the group of heat shockproteins from alfalfa and heat shock protein hydrolysates from alfalfa,the product further comprising (ii) a drug suitable for treating a skindisorder.

In particular, the invention relates to a healthcare product comprising(i) a component selected from the group of heat shock proteins fromalfalfa and heat shock protein hydrolysates from alfalfa, the productfurther comprising imiquimod or pharmaceutical acceptable salt orderivative thereof.

Imiquimod is a pharmaceutical compound represented by the followingformula.

The imiquimod may be provided as the free base or in a pharmaceuticallyactive salt-form or a pharmaceutically active derivative form. As usedherein a derivative means in particular a molecule formed from thereaction of the

—NH₂ group of imiquimod with another compound, in particular a compoundforming a hydrolysable group, such as an amide, with imiquimod.

The term “or” as used herein is defined as “and/or” unless specifiedotherwise.

The term “a” or “an” as used herein is defined as “at least one” unlessspecified otherwise.

When referring to a moiety (e.g. a compound) in the singular, the pluralis meant to be included, unless specified otherwise.

When referring to a ‘drug’ or ‘pharmaceutical compound’, this does NOTinclude Hsp or Hsp hydrolysate, but to a different compound having apharmaceutical activity in the treatment of the skin. Generally thepharmaceutical compound is a relatively small organic compound, having amolecular mass of less than 1000 g/mol. Specifically the term is usedfor imiquimod in all its pharmaceutically active and pharmacologicallyacceptable forms.

A healthcare product or Hsp containing composition according to theinvention is in particular useful for the prophylactic or therapeutictreatment of a skin disorder.

In particular, the inventors have found that the use of Hsp inco-therapy with a drug applied for prophylactic or therapeutic treatmentof a skin disorder is advantageous in the prophylaxis or therapeutictreatment of a negative side effect of the drug, more specifically inthe prophylaxis or therapeutic treatment of a negative side effect ofimiquimod. Possible side effects of imiquimod include eczema, alopecia,dermatitis, and pruritus. Remote site skin reaction has been reportedand included erythema (3% in females, 1% in males), ulceration (2% infemales), erosion (2% in males), edema (1% in females, 1% in males),induration (1% in males), and scaling/excoriation/flaking (1% in males).Remote site reactions reported in more than 1% of patients have includedbleeding, burning, itching, pain, dryness, scabbing, crusting, redness,hardening of the skin, tenderness, and tinea cruris. Hyperkeratosis,rash, skin disorder, photosensitivity reaction, verruca, and remote siteirritation have been reported. Imiquimod-induced psoriasis and at least1 case each of vitiligo-like hyperpigmentation, contact pemphigus,pityriasis rubra pilaris exacerbation, erosive pustular dermatosis ofthe scalp, and eruptive keratoacanthoma have been reported. Exfoliativedermatitis, erythema multiforme, hyperpigmentation, and hypertrophicscar have been reported during postmarketing experience.

Further side-effects include change of color of treated skin, headache,dizziness, chest pain, back pain, cold sores, fever blisters, coldsymptoms such as stuffy nose, sneezing, sore throat, nausea, diarrhea,loss of appetite, vaginal itching or discharge, Arthritis.

In particular, it has been found that the application of Hsp is usefulfor avoiding or reducing the occurrence of erythema, scaling of theskin, or thickening of the skin.

Accordingly, the invention further relates to a heat shock protein, foruse in preventing the occurrence of a negative side effect of atreatment with imiquimod or another drug for treating a skin disorder,or therapeutically treating such side effect. The invention has beenfound particularly effective for the prophylaxis of a negative sideeffect of imiquimod, such as scaling, or at least to reduce the severityat which a side-effect of imiquimod or another drug manifests itself.

That the product is effective in a prophylactic treatment can beverified routinely by comparing the incidence of the negative effect ina first population (e.g. a test panel of humans, or in test-animals)treated with the drug but not the Hsp and a second population(comparable with the first) treated with both the drug and the Hsp.

A product of the invention may in particular be for use in the treatmentof mammal, preferably a human. The human may be an infant (<1 yr), anolder child (1-12 yr), an adolescent (12-18 yr), or an adult in hisearly (19-39 yr), middle (40-65 yr) or late adulthood (>65 yr).

A product according to the invention may in particular be suitable forpeople or animals who have an impaired production of endogenous Hsp, orwho are not capable of producing endogenous Hsps at all. It may forexample be suitable for elderly people (more than 50, more than 60 ormore than 70 years of age) or for people having scar tissue in or ontheir skin.

The production of an endogenous Hsp in skin of a subject is inparticular considered impaired, if the start of the endogenous Hspproduction after exposure of skin cells of the subject to a trigger forinducing endogenous Hsp is delayed (i.e. an increased response time)and/or if the endogenous Hsp production (per hour) is reduced, afterexposure to the trigger. A particularly suitable trigger is exposure ofthe skin cells to a temperature of 43° C. (as described in U.S. Pat. No.6,737,086).

The skin cells (keratinocytes and/or fibroblasts) can be taken from thesubject and incubated in a manner known in the art per se. Hsp levelscan be determined as described in S. Sanchez et al., Radiation Research167 (2007) 572-580.

By comparing produced amount of endogenous Hsp and/or response time witha normal reference value (from cells with normal Hsp production,preferably from a subject of about the same age, the same sex and thesame or a similar skin type), it can be established whether endogenousHsp production is impaired. The produced amount can suitably bedetermined after 1 hour, after 2 hours, after three hours or after alonger exposure, depending on the minimum response time that isconsidered normal for a specific subject. For example, in U.S. Pat. No.6,737,086 it was reported that in human skin cells originating from afive-year-old subject Hsp90 production occurred after 1 hour of exposureto a temperature of 43° C.

In impaired skin cells, the response time may in particular be increasedby a factor of at least 1.5, at least 3, or at least 6. In impaired skincells, the total amount of produced endogenous Hsp (after 1 hr, after 2hrs or after 3 hrs) may in particular be reduced by a factor of at least1.5, at least 3, or at least 6.

A product according to the invention may in particular be used to treata skin disorder selected from the group of acne (for exampleadolescents), warts, athlete's foot, Lyme disease, psoriasis, lichen,ichthyosis, keratosis, Darier's disease, pustulosis, herpes (inparticular herpes zoster), cellulitis, eczema (such as atopicdermatitis), neurodermatitis (such as lichen simplex chronicus, prurigonodularis, lichen striatus or atopic dermatitis), inflammatory skindisorders and children's diseases affecting the skin (such as varicella,rubella, measles).

In an embodiment, the product is for use in the prophylactic ortherapeutic treatment of a disorder selected from the group of molluscumcontagiosum, vulvar intraepithelial neoplasia, and vaginalintraepithelial neoplasia.

In an embodiment, the product is for use in the prophylactic ortherapeutic treatment of a disorder selected from the group ofSuperficial and Nodular BCC, Sclerodermiform BCC, Bowens disease,invasive squamous cell carcinoma, Lentigo Maligna, Metastatic Melanoma,Mycosis Fungoides, Keratoacanthoma, Extramammary Pagets Disease.

In an embodiment, the healthcare product or Hsp (hydrolysate) is for usein the prophylactic or therapeutic treatment of psoriasis. In a specificembodiment, the psoriasis is imiquimod-induced psoriasis.

The healthcare product according to the invention may contain a singlecomposition for administration to an intended user comprising both Hsp(or hydrolysate thereof) and drug (imiquimod) or it may comprise two ormore separately packaged compositions for administration to the intendeduser. Thus, the Hsp (hydrolysate) and the drug for skin treatment maybut do not need to be present in the same composition.

The healthcare product usually is a skincare product, i.e. it comprisesat least one component that is intended to be applied to the skin.Typically, at least the pharmaceutical compound for skin treatment, suchas imiquimod, is present in a skincare composition for application tothe skin. The Hsp (hydrolysate) may be present in the same skincarecomposition or a different skincare composition. However, it also may beadministered in a different manner, notably orally or nasally.

Good results have been achieved with a product wherein Hsp and the drugfor skin treatment (imiquimod) have been applied separately.

Accordingly, in an advantageous embodiment, the healthcare productcontains a first container containing a first healthcare compositioncomprising the heat shock protein or heat shock protein hydrolysate anda second container containing a second healthcare composition comprisingimiquimod.

A healthcare product, first composition or second composition that isfor application to the skin may in particular be selected from the groupof creams, lotions, powders, gels, foams, oils, sprays (e.g. aerosolsprays), mousses, salves, and balms.

In an advantageous embodiment, the first composition is foradministration to the skin.

In particular, in case a fast response is desired, it is considered toprovide a first composition for administration via the respiratorytract, preferably a composition selected from nasal sprays, compositionfor oral administration into the respiratory tract, such as an inhaler.

Alternative administration forms for the Hsp (hydrolysate) includecompositions for oral or rectal intake into the gastro-intestinal tractor compositions for mucosal application.

In yet another embodiment, the Hsp is in a form for parenteraladministration. Thus Hsp may be in a form for intravenous,intramuscular, intracutaneous or subcutaneous injection.

Generally, the Hsp or hydrolysate thereof that has been separated, inparticular isolated from the source (the plant or part thereof) whereinit has been produced.

The Hsp is usually obtained from a plant, e.g. from a fluid from a plantor from a plant extract. Suitable methods to obtain the Hsp are known inthe art per se, for instance from EP 1 531 160 A1.

The plant Hsp may be provided in a manner known per se. For instance,the plant Hsp may be a plant Hsp obtained by a method described in EP-A1531160, of which the contents—in particular the claims and examples—areincorporated herein by reference.

In particular the Hsp may be a plant Hsp obtained by a method accordingto WO 2010/115990, of which the contents—in particular the examples,suitable process conditions and suitable materials for use in themethod—are incorporated herein by reference.

Preferably, the Hsp is a natural, i.e. non-recombinant, Hsp; theHsp-hydrolysate preferably is a hydrolysate from natural(non-recombinant) Hsp. It is contemplated that a natural Hsp orhydrolysate of natural Hsp may be tolerated better by the subjecttreated with the skin-care product, in particular that the risk ofallergenic reactions may be less. Further, consumer acceptance may bebetter for non-recombinant plant Hsp.

A product according to the invention may comprise native Hsp ordenaturated Hsp.

A product according to the invention may in particular comprise heatshock proteins or heat shock protein hydrolysates from alfalfa (Medicagosaliva).

Optionally, in addition or alternatively the skin care product comprisesHsp or Hsp-hydrolysate from another plant. Other plants as a source ofHsp may in particular be selected from the group of cereals (forinstance barley), soy, grasses (for instance oat), peas, beet, potato,clover and water plants (for instance an alga).

In particular, leaves of the plant may be used as source for one or moreHsps. Particularly suitable are beet tops, alfalfa leaves, barleyleaves, oat leaves and potato tops.

With Hsp-hydrolysate is meant Hsp wherein part of the chemical bondshave been hydrolysed, in particular peptide bonds. Generally, forachieving an intended cosmetic or medical effect, non-hydrolysed Hsp maybe particularly suitable. It is however contemplated that in someembodiments, the presence of Hsp-hydrolysate may be advantageous. Forexample, it is contemplated that hydrolysed Hsp may penetrate betterinto (the cells of) the skin.

The Hsp-hydrolysate may be prepared by, e.g., chemical hydrolysis,enzymatic hydrolysis or a combination thereof. Chemical hydrolysis mayfor example be performed in an aqueous medium of neutral pH, or in anaqueous medium in the presence of an acid (e.g. a strong acid such asHCl) or a base (e.g. a strong base such as NaOH). The hydrolysis(chemical and/or enzymatic) may be carried out at an elevatedtemperature. The enzymatic hydrolysis may in particular be performedwith a proteolytic enzyme, based on technology known per se. Enzymatichydrolysis is an effective alternative to chemical hydrolysis, becauseit is relatively mild in comparison to acid or alkali hydrolysis.Additionally, the inherent specificity of a specific proteolytic enzymeof choice can control the nature and extent of hydrolysis, and thus thefunctional properties of the end product.

The degree of hydrolysis may be chosen within wide limits. At least 10wt. %, at least 25 wt. %, at least 50 wt. %, at least 80 wt. % or atleast 90 wt. % (based on the sum of Hsp-fragments and unhydrolysed Hsp)of the Hsp-hydrolysate may be formed by Hsp fragments. Of theHsp-hydrolysate, 100 wt. % or less, 95 wt. % or less, at least 75 wt. %or less, 50 wt. % or less or 25 wt. % or less (based on the sum ofHsp-fragments and unhydrolysed Hsp) may be formed by Hsp fragments.

The size of the fragments may be chosen within wide limits. Usually, incase a hydrolysate is present, at least 50 wt. %, in particular at least75 wt. %, more in particular at least 90 wt. % (based on the sum ofHsp-fragments and unhydrolysed Hsp) of the hydrolysate is formed bypeptides (including unhydrolysed Hsp) having at least five amino acidresidues. In a specific embodiment, at least 25 wt. %, in particular atleast 50 wt. %, more in particular at least 75 wt. % (based on the sumof Hsp-fragments and unhydrolysed Hsp) of the hydrolysate is formed bypeptides (including unhydrolysed Hsp) having at least ten amino acidresidues.

The skin-care product may in particular comprise at least one Hsp orhydrolysate thereof selected from the group of Hsp40, Hsp60, Hsp70 andHsp90, respectively hydrolysates of any of these Hsps. In particular,Hsp70 or a hydrolysate thereof is preferred.

In a specific embodiment, Hsp40 or hydrolysate thereof is present. Hsp40is expected to act as a co-chaperone and may thus improve the efficiencyof Hsp70, for example by increasing the ATPase activity of Hsp70.

The concentration of Hsp, Hsp hydrolysate, or a mixture thereof in aproduct according to the invention may be chosen within wide limits,usually in the range of 0.1 μg to 1 mg per mL of the composition inwhich it is provided (i.e. the skin care product if it is formed of asingle composition or in case of a product comprising more than onephysically separated compositions, said first composition). Inparticular, the Hsp concentration is at least 5 μg/mL, preferably atleast 20 μg/mL. In particular good results have been achieved with aconcentration of about 50 μg/mL or more, more in particular about 0.2mg/ml or more. Preferably the Hsp concentration is about 0.6 mg/mL orless, in particular about 0.5 mg/mL or less.

The concentration of imiquimod (or another drug for treatment of a skindisorder, usually is at least 0.001 mg/ml of the composition in which itis provided (i.e. the skin care product if it is formed of a singlecomposition or in case of a product comprising more than one physicallyseparated compositions, said second composition). Preferably, theconcentration is at least 0.01 mg/ml, in particular 0.02 mg/ml or more.Usually, the concentration is 10 mg/ml or less, preferably 1 mg/ml orless in particular 0.5 mg/ml or less, more in particular 0.1 mg/ml orless.

The healthcare product, the first composition or the second composition,may further comprise known ingredients for a specific type of healthcareproduct, or composition, e.g. for a lip balm, for a moisturizing cream,for a body lotion, for a massage oil, for a scrub crème, for a peelingcrème, for a depilatory cream. Suitable examples of known ingredientsare for example UV blocking agents, preservatives, stabilizers,moisturizers, antioxidants, vitamins, fragrances, thickening agents,chalk, ceramides, emulsifiers, surfactants, minerals, alkaloids,enzymes, co-enzymes, acids, polyphenols, ceramides, herbs, plantextracts, solvents, amino acids, oil lipids, pH adjuster, salts,polysaccharides, fatty acids, flavonoids, hormones, yeast extracts,matrix metalloproteinases, peptides, emoillents.

A skin care product, the first composition or the second composition maybe based on commercially available or otherwise known formulations towhich the drug (imiquimod) or Hsp or Hsp-hydrolysate is added, e.g. on aknown salve, (lip) balm, lotion, mousse, cream, oil, powder, gel, foam,spray. Examples of known skin care compositions are for example given inWO 01/85129 A2.

In an embodiment, the ingredients of the skin care product/composition,including the Hsp or Hsp hydrolysate (if present) or the drug (ifpresent), are dissolved or emulsified in a lipophilic medium, which maycomprise one or more components selected from the group oftriglycerides, such as capric triglyceride, C18-C36 alkyl acidtriglycerides; acrylates, such as C10-C30 alkyl acrylates; oils, such asolive oil, sunflower oil, sclerocarya birrea oil, lime oil, nut oil,manuka oil, mineral oil, rapeseed oil, teatree oil; hydroxyethyl urea;isodecyl laurate; fatty acids such as stearic acid; and phospholipidssuch as lecithine.

In particular, in a spray for application to the skin, the ingredientsmay be suspended or emulsified in a propellant, which may comprise oneor more components selected from the group of chlorofluorocarbons suchas 1,1,1,2-tetrafluoroethane, trichlorotrifluoroethane; hydrocarbongases such as propane, isobutane and isopentane; dimethyl ether; carbondioxide; and nitrogen gas.

In addition to the Hsp or Hsp-hydrolysate, a product according to theinvention may comprise one or more other active agents, e.g. aco-chaperone for the Hsp or Hsp-hydrolysate. However, the product mayalso be free of such additional agents. It is contemplated that the Hspor Hsp-hydrolysate may be capable of interacting with an endogenous(formed in situ by the subject) co-chaperone or the like.

In addition, one or more preserving agents may be present such assorbate, benzoate, sulfite, or the like.

As will be understood by the skilled person, the composition isformulated to be safe for its intended administration.

The drug (imiquimod), Hsp or Hsp hydrolysate in a product according tothe invention may be incorporated in a carrier, preferably acosmetically or pharmaceutically acceptable carrier. It may for examplebe incorporated in a liposome or a microcapsule.

In the context of the invention liposomes are defined as compositestructures comprising lipids, in particular phospholipids, and maycontain small amounts of other molecules. The sizes of liposomes areusually in the range of 20 nm to 1000 nm. They may for example be atleast 40 nm, at least 100 nm or at least 250 nm. The liposomes may forexample be 800 nm or less, 600 nm or less or 400 nm or less.

It is envisaged that a carrier, in particular a liposome that comprisesthe drug (imiquimod), Hsp or a hydrolysate thereof may be capable ofeffective delivery of the drug (imiquimod) an Hsp respectively ahydrolysate thereof into cells of the skin.

It is further envisaged that incorporation of the drug (imiquimod) Hspand/or a hydrolysate thereof in a carrier may result in an increasedstability of the drug (imiquimod), Hsp respectively the hydrolysatethereof, for example because of a decreased exposure to (atmospheric)oxygen.

It is contemplated that at least in some embodiments Hsp or Hsphydrolysate may be negatively affected by an alcohol, for instanceethanol. Accordingly, it is advantageous to use no alcohol or only as aminor component (e.g. in aqueous liquid) during the recovery process ofthe Hsp from the source and during the preparation of the skin-careproduct.

Imiquimod or another drug for treating a skin disorder,The concentration of the drug, in particular imiquimod, is usually inthe range of 0.1-10 wt. %, preferably in the range of 0.5-8 wt. %. Inparticular for imiquimod, the concentration is preferably in the rangeof 1-6 wt. %. Typical examples of formulations are a 5 wt. % lotion orcream, a 3.75 wt. % lotion or cream and a 2.5 wt. % lotion or cream.

The invention further relates to a method for the prophylactic treatmentof a skin disorder. The invention further relates to the prophylaxis ofa negative side-effect of a healthcare product comprising a therapeuticcompound for the treatment of a skin disorder. The invention furtherrelates to a method for the therapeutic treatment of a skin disorder.The invention further relates to the therapeutic treatment of a negativeside-effect of a healthcare product comprising a therapeutic compoundfor the treatment of a skin disorder.

The skin disorder or side-effect in a method according to the inventionis preferably selected from those mentioned herein above. Preferredproducts for use in the method are those as described elsewhere herein.

A method of treatment according to the invention typically comprisesadministering an effective dose of Hsp or Hsp hydrolysate to a subject,preferably a human. The human is generally treated with a drug fortreating a skin disorder, preferably imiquimod, has been treated with adrug for treating a skin disorder, preferably imiquimod, or it isintended that the subject is to be treated with a drug for treating askin disorder, preferably imiquimod.

Imiquimod or the other drug is typically applied to the skin. It istypically applied before, after or together with the Hsp or Hsphydrolysate.

In a practical embodiment, said administration of Hsp or Hsp hydrolysateis a topical administration of a composition comprising Hsp or Hsphydrolysate to a part of the skin that is/has been or is intended to betreated with imiquimod or another drug.

The treatment by administration on the skin can suitable done by e.g.spraying onto the skin, applying onto the skin by an applicator, such asa roller, a brush or a sponge, or by manual application.

In a further embodiment, the Hsp or Hsp hydrolysate is administered viathe respiratory tract, nasally or orally. An advantage thereof is fastway of action. Nasally given Hsp has been shown to be present in musclesand is able to pass the blood brain barrier.

In a further embodiment the Hsp is administered into thegastro-intestinal tract, typically orally or rectally.

In a further embodiment, the Hsp is administered parentally(intravenous, intramuscular, intracutaneous or subcutaneous injection).An advantage thereof is a fast way of action.

The dosage of Hsp or hydrolysate thereof may be chosen within widelimits, depending on the intended use, the subject and the way ofadministration. As a rule of thumb a suitable average daily dosage ischosen in the range of 0.1 μg and 1 mg per square centimetre of treatedskin. Usually, the average daily dosage is 0.5 μg per square centimetreof treated skin or more, in particular 1 μg per square centimetre oftreated skin or more. Usually, the average daily dosage is 500 μg persquare centimeter of treated skin or less, in particular 250 μg persquare centimetre of treated skin or less.

In a further embodiment, in particular in an embodiment where the Hsp isnot administered topically to the skin, the dosage may be in the rangeof 0.1 μg-25 mg/kg body weight, in particular in the range of 0.5 μg-15mg/kg body weight, more in particular in the range of 2 μg-10 mg/kg bodyweight.

The dosage of imiquimod or other drug for treating a skin disorderthereof may be chosen within wide limits, depending on the intended use,the subject and the way of administration. As a rule of thumb a suitableaverage daily dosage is chosen in the range of 0.1μg and 5 mg per squarecentimetre of treated skin. Usually, the average daily dosage is 1 μgper square centimetre of treated skin or more, in particular 50 μg persquare centimetre of treated skin or more. Usually, the average dailydosage is 4 mg per square centimetre of treated skin or less, inparticular 2 mg per square centimetre of treated skin or less.

The skin care product may be administered e.g. once a week; preferablythe skin care product is administered at least once a day. The productmay be administered a plurality of times per day, e.g. 2-10 times, 2-6times or 2-3 times. In case the product comprises more than onecompositions (such as the first composition comprising the Hsp(hydrolysate) or the second composition comprising the drug, such asimiquimod), the compositions do not need to be administered the samenumber of times per week or day.

Good results have been achieved with a method wherein first a firstcomposition comprising Hsp or Hsp hydrolysate is administered, andwherein thereafter a second composition comprising the imiquimod orother drug is administered.

Good results have been achieved with a method wherein there is atime-interval between administration in order to allow the Hsp to beincluded in the skin. The interval may in particular be chosen between 1min and 8 hours, more in particular between 30 min and 6 hours, more inparticular between 1 hour and 4 hours.

In a practical embodiment, Hsp is applied at the same time as imiquimodor the other drug.

In an advantageous embodiment, another dosage of a compositioncomprising Hsp or Hsp hydrolysate is given after administration of thecomposition comprising imiquimod or other drug. The interval between thesubsequent dosage of Hsp (hydrolysate) and imiquimod or other drug canbe chosen in the same range as for the interval between the Hsp(hydrolysate) dosage preceding the administration of the drug and saidadministration.

A dosage regime as described above advantageously repeated at least oncea day.

The product or a component thereof (such as the first compositioncomprising the Hsp (hydrolysate) or the second composition comprisingthe drug, such as imiquimod) may be present in or on a plaster that isapplied on at least a part of the skin that is in need of treatment.Advantageously, such plaster gradually releases the amount of skin careproduct that is intended to administered, for example in 24 hours orless, in 12 hours or less, in 6 hours or less or in 3 hours or less.

The invention will be illustrated by the following example.

EXAMPLE: TOPICAL ALFALFA-DERIVED HSP70 FOR TREATMENT OF A SKIN DISORDER

Materials

Imiquimod was provided in a commercially available cream (Fougera, acream comprising 5% Imiquimod).

HSP70 (from alfalfa) was obtained from Alfa Biogene International B.V.It was dispersed in a concentration of 50 or 250 μg/ml in a carriercream from Deutscher Apotheken Index. Said carrier cream contained (per100 g):

-   -   4.0 g Glycerolmonostearate 60    -   6.0 g Cetylalcohol    -   7.5. g Medium-chain Triglycerides    -   25.5 g White Petrolatum    -   7.0 g Macrogol-20-glycerolmonostearate    -   10.0 g Propylene-glycol    -   40.0 g Purified water

Methods

The effect of HSP was tested on mice (BALBc mice, male 10-12 weeks old)

The mice were divided in 5 groups of eight mice each:

Group 1: mice were treated for 6 days on shaved back and right ear withone daily application of imiquimod 5% cream to the skin (3.125 mg dailydose)

Group 2: mice were treated for 6 days as group 1. In addition the partof the skin to which imiquimod was applied was treated twice daily withthe cream comprising 50 μg/ml HSP (20 μg HSP per dose). Each day, thecream comprising HSP70 was applied to the skin in a 1 mm layer, 4 hoursbefore application of the imiquimod, and again 4 hours after treatmentwith imiquimod.

Group 3: mice were treated for 6 days as group 2, except for the creamcomprising 250 μg/ml HSP (100 μg HSP per dose)

Group 4: mice were treated as group 2 except for the carrier creamcomprising no Hsp70. (mice were treated for six days, twice a day, oncefour hours before treatment with imiquimod, once four hours aftertreatment with imiquimod.

Group 5: control group, no cream application.

Study Protocol:

Day 1: initial evaluation for visible signs of psoriasis. All visiblesigns were categorized and scored by a PASI score (PASI: psoriasis areaand severity index) from 0-4 (erythema, scales, thickness, cumulativescore: maximum of 12 points). Animal backs were shaved and photographswere taken.

Thickness of both ear pinnae were measured. Cream application wasperformed

according to the categorization into above detailed groups.

Day 2-6: Animals in the treatment groups were treated with the accordingskin

creams. All animals were assessed and examined once daily forbehavioural changes and adverse effects. Ear pinnae measurements areperformed daily. Skin changeswere assessed by the PASI score. On day 7, photographs of all animals(backs) were taken to document progression of the disease.

Day 7: The animals were euthanized and skin tissue and blood wasprocessed. Inflammatory markers were measured with Q-PCR (skin samples).Skin samples were histologically examined.

Statistical Methods:

Study groups on ordinal and continuous outcomes were compared using theKruskal-Wallis test with a Steel-Dwass adjustment for pairwise groupcomparisons for an overall statistical significance criteria of 0.05 foreach outcome.

Results

Ear Pinnae:

Thickness of the right ear remained in the range of 0.20-0.25 mm foreach of the groups until day 4. Thereafter, the thickness rose sharplyfor Group treated with imiquimod (Group 1) only, and to a less extentfor the other Groups (2-4). The following table shows ear thickness inmm.

Group Day 1 Day 5 Day 6 Day 7 1 (imiquimod only) 0.21 0.28 0.37 0.40 2(20 μg HSP) 0.23 0.28 0.33 0.34 3 (100 μg HSP) 0.22 0.25 0.28 0.29 4(carrier cream) 0.22 0.28 0.30 0.33 5 (control) 0.25 0.22 0.19 0.19

The results show that HSP used in co-therapy with imiquimod is effectivein reducing or even avoiding the occurrence of a side-effect ofimiquimod (swelling).

Epidermal Thickness:

A histology study on the back skin samples taken of the euthanized mice,resulted in the following average epidermal thickness of the treatedskin (in μm):

Group epidermal thickness 1 (imiquimod only) 58 2 (20 μg HSP) 42 3 (100μg HSP) 40 4 (carrier cream) 53 5 (control) 10

The results show that HSP used in co-therapy with imiquimod is effectivein reducing or even avoiding the occurrence of a side-effect ofimiquimod (swelling).

PASI-Score:

The results for the cumulative PASI Score on day 7 were as follows:

Group PASI score 1 (imiquimod only) 6.5 2 (20 μg HSP) 1.7 3 (100 μg HSP)3.0 4 (carrier cream) 3.5 5 (control) 0

The results support that HSP can be used to reduce psoriasis area andseverity.

Visual Appearance:

FIG. 1 shows the backs of one mouse of each of groups 1-4 after 6 daysof treatment. From left to right: Group 2 (20 μg HSP), Group 3 (100 μgHSP), Group 4 (carrier cream only) and Group 1 (imiquimod only)

As illustrated by FIG. 1, the backs of the mice treated with imiquimodin co-therapy with HSP were still essentially free of scaling, whereasthe mice treated with imiquimod only were severely scaled. Increasedscaling was also observed in mice treated with carrier cream only,compared to the mice treated in accordance with the invention.

1.-10. (canceled)
 11. A method for the prophylactic or therapeutictreatment of a skin disorder selected from the group consisting of acne,warts, athlete's foot, Lyme disease, psoriasis, lichen, ichthyosis,keratosis, Darier's disease, pustulosis, herpes zoster, cellulitis,eczema, neurodermatitis, herpes, inflammatory skin disorders, skincancers, and children's diseases affecting the skin comprisingadministering to a patient in need thereof a healthcare productcomprising (i) a component selected from the group consisting of heatshock proteins from alfalfa and heat shock protein hydrolysates fromalfalfa, the product further comprising Imiquimod or a pharmaceuticallyacceptable salt or derivative thereof.
 12. The method of claim 11,wherein the skin disorder is psoriasis.
 13. The method of claim 11,wherein the skin disorder is a skin cancer selected from the groupconsisting of basal cell carcinoma, Bowen's disease, superficialsquamous cell carcinoma, superficial malignant melanomas, and actinickeratosis.
 14. The method of claim 11, wherein the skin disorder iswarts.
 15. A method of alleviating a negative-side effect of imiquimodcomprising administering to an individual in need thereof imiquimod anda heat shock protein (Hsp).
 16. The method of claim 15, wherein thenegative side effect is selected from the group consisting of erythema,scaling of the skin, and thickening of the skin.
 17. The method of claim15, wherein the Hsp is included in a first healthcare composition andthe imiquimod is included in a second healthcare composition and whereinthe first healthcare composition and the second healthcare compositionare administered, and wherein at least the second healthcare compositionis applied to the skin.
 18. The method of claim 15, wherein the Hsp andthe imiquimod are administered simultaneously.
 19. The method of claim15 wherein the heat shock protein is Hsp
 70. 20. The method of claim 15wherein the heat shock protein is Hsp 70 from alfalfa.